Impact of heat-induced retrieval methods on silver staining for detecting senile plaques of Alzheimer's disease in human brain tissue sections.
8 affiliations
- 1Department of Anatomy, School of Basic Medical Sciences, Anhui Medical University, Hefei, Anhui, 230032, P. R. China.
- 2Department of Pharmacy, the First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230022, P. R. China.
- 3School of Pharmacy, Anhui Medical University, Hefei, Anhui, 230032, P. R. China.
- 4Institute of Brain Science, the First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230022, P. R. China.
- 5Department of Pharmacy, the First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230022, P. R. China. [email protected].
- 6School of Pharmacy, Anhui Medical University, Hefei, Anhui, 230032, P. R. China. [email protected].
- 7Department of Anatomy, School of Basic Medical Sciences, Anhui Medical University, Hefei, Anhui, 230032, P. R. China. [email protected].
- 8Institute of Brain Science, the First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230022, P. R. China. [email protected].
Abstract
Background
Silver staining continues to serve as a critical technique for histological detection of pathological deposits, especially in the context of Alzheimer's disease (AD). In clinical research, paraffin-embedded sections are often stored for extended periods, and tissues may remain in formaldehyde for prolonged durations, both of which can lead to false-negative results. The objective of this study was to systematically evaluate whether heat-induced retrieval can enhance the efficacy and sensitivity of classical silver staining in these suboptimally preserved brain tissues.Methods
This study investigated the impact of heat-induced retrieval methods on silver staining efficacy in brain tissue. Three types of samples were examined: standard FFPE sections, tissues immersed in formaldehyde for 48 months, and paraffin‑embedded sections (initially fixed for 48 h in formalin, sections coated with paraffin and stored on slides) stored for seven years. Retrieval methods were performed using citrate-citric acid buffer, Tris-EDTA buffer, or proteinase K. Silver staining was conducted using three classical methods: Bielschowsky, periodic acid-methenamine silver (PAM), and Gallyas; untreated sections were used as controls.Results
Results showed that both citrate-citric acid and Tris-EDTA significantly increased the signal-to-noise ratio of senile plaques in Bielschowsky staining. This improvement facilitated better the detection of amyloid plaque details and density, with Tris-EDTA yielding higher sensitivity in long-stored paraffin sections. Proteinase K digestion did not enhance Bielschowsky staining results. None of the retrieval methods improved signal-to-noise ratio for senile plaques or neurofibrillary tangles with PAM or Gallyas.Conclusions
These findings indicate that heat-induced retrieval methods with citrate or Tris-EDTA buffer, enabling more efficient silver ion reduction at specific targets in Bielschowsky. This work provides an experimental basis for optimizing silver staining protocols, which helps enhance diagnostic accuracy in neuropathological studies of AD and related disorders.
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BibTeX
@article{zhang2026impact,
title = {{Impact of heat-induced retrieval methods on silver staining for detecting senile plaques of Alzheimer's disease in human brain tissue sections.}},
author = {Zhi-Yi Zhang and Shan-Shan Zhang and Shuai-Deng Wang and Kun Cheng and Wen-Hui Yi and Zi-Wei Li and Jiang-Ning Zhou and Xin-Meng Wang and Chen-Wei Wang},
journal = {Diagnostic pathology},
year = {2026},
month = jul,
doi = {10.1186/s13000-026-01799-3},
url = {https://europepmc.org/article/MED/42968096},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Impact of heat-induced retrieval methods on silver staining for detecting senile plaques of Alzheimer's disease in human brain tissue sections.
AU - Zhi-Yi Zhang
AU - Shan-Shan Zhang
AU - Shuai-Deng Wang
AU - Kun Cheng
AU - Wen-Hui Yi
AU - Zi-Wei Li
AU - Jiang-Ning Zhou
AU - Xin-Meng Wang
AU - Chen-Wei Wang
PY - 2026
DA - 2026/07/29
JO - Diagnostic pathology
DO - 10.1186/s13000-026-01799-3
UR - https://europepmc.org/article/MED/42968096
AB - Background Silver staining continues to serve as a critical technique for histological detection of pathological deposits, especially in the context of Alzheimer's disease (AD). In clinical research, paraffin-embedded sections are often stored for extended periods, and tissues may remain in formaldehyde for prolonged durations, both of which can lead to false-negative results. The objective of this study was to systematically evaluate whether heat-induced retrieval can enhance the efficacy and sensitivity of classical silver staining in these suboptimally preserved brain tissues.Methods This study investigated the impact of heat-induced retrieval methods on silver staining efficacy in brain tissue. Three types of samples were examined: standard FFPE sections, tissues immersed in formaldehyde for 48 months, and paraffin‑embedded sections (initially fixed for 48 h in formalin, sections coated with paraffin and stored on slides) stored for seven years. Retrieval methods were performed using citrate-citric acid buffer, Tris-EDTA buffer, or proteinase K. Silver staining was conducted using three classical methods: Bielschowsky, periodic acid-methenamine silver (PAM), and Gallyas; untreated sections were used as controls.Results Results showed that both citrate-citric acid and Tris-EDTA significantly increased the signal-to-noise ratio of senile plaques in Bielschowsky staining. This improvement facilitated better the detection of amyloid plaque details and density, with Tris-EDTA yielding higher sensitivity in long-stored paraffin sections. Proteinase K digestion did not enhance Bielschowsky staining results. None of the retrieval methods improved signal-to-noise ratio for senile plaques or neurofibrillary tangles with PAM or Gallyas.Conclusions These findings indicate that heat-induced retrieval methods with citrate or Tris-EDTA buffer, enabling more efficient silver ion reduction at specific targets in Bielschowsky. This work provides an experimental basis for optimizing silver staining protocols, which helps enhance diagnostic accuracy in neuropathological studies of AD and related disorders.
SN - 1746-1596
ER -